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Virologica Sinica, 19 (4) : 320-324, 2004
Research Article
Study of Cell Culture System Transfected with Full-length cDNA Clone of Hepatitis C Virus*
1. College of Life Scinces, Wuhan University, Wuhan 430072, China
2. Chinawave Biotechnology Co. Ltd, Guangzhou 510010, China
A Vero-E6 culture system was developed to assay Hepatitis C virus (HCV) replication by plasmid (pHCV) transfection,which contains a T7 promoter at the 5’end, full-length cDNA of the HCV genome and a T7 terminator. To facilitate high-level transcription of HCV RNA, Vero-E6 cells were transfected with pHCV and then infected with recombinant vaccinia virus (vTF7-3) containing the T7 RNA polymerase gene. This transfection-based cell culture system produced high levels of HCV genome (107-108 copies/mL) detectable by real-time quantitative PCR and the production of HCV RNA transcripts was confir med by RT-PCR. Western blot analysis revealed that HCV structural and non-structural proteins were correctly processed. The transfected Vero-E6 cells assembled 40-50nm virus-like particles were analysed with transmission electron microscope. This model can be utilized for studying mechanisms of HCV replication, preparation of HCV vaccine and to test potential antiviral drugs.
  Published online: 25 Aug 2004
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